[Super SMART cDNA synthesis technology for amplifying small amount of total RNA of peripheral blood eosinophils from asthma patients]

Hai-jin Zhao, Shao-xi Cai, Fei Zou, Wan-cheng Tong, Wei-ren Wan
Di 1 Jun Yi da Xue Xue Bao, Academic Journal of the First Medical College of PLA 2004, 24 (9): 1037-9

OBJECTIVE: To amplify double-strand cDNA from small amount of total RNA of eosinophils from asthma patients by Super SMART cDNA synthesis technique.

METHODS: The eosinophils were purified from the peripheral blood of asthma patients before and after treatment by Percoll gradient centrifugation, from which the total RNA was extracted using TRIzol kit. First-strand cDNA synthesis and double-strand cDNA amplification were performed using Super SMART cDNA synthesis technique. The quality of the obtained cDNA was evaluated by gradient cDNA electrophoresis, and the amplification efficiency determined by cDNA quantification.

RESULT: From 20 ng total RNA, 7.155 microg and 6.568 microg of the tester and driver double- strand cDNAs respectively were obtained successfully, and the result of electrophoresis indicated high quality and purity of the cDNA acquired.

CONCLUSION: Super SMART cDNA synthesis technique can effectively amplify high-quality double-strand DNA from a very small amount of total RNA, which may facilitate the exploration of the mechanism of asthma from the genetic level.

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